LOCUS       sequence-415151-        8748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Premade nuclei membrane marker, pUBQ10:H2B:PmScarlet-I:tHSP18.2M, 
            for braiding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8748)
  AUTHORS   Piepers M, Erbstein K, Reyes-Hernandez J, Song C, Tessi T, 
            Petrasiunaite V, Faerber N, Distel K, Maizel A
  TITLE     GreenGate 2.0: Backwards compatible addons for assembly of complex 
            transcriptional units and their stacking with GreenGate.
  JOURNAL   PLoS One. 2023 Sep 8;18(9):e0290097. doi: 
            10.1371/journal.pone.0290097. eCollection 2023.
  PUBMED    37682951
REFERENCE   2  (bases 1 to 8748)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2023-09-8"; volume: "18(9):e0290097. doi"; pages: " 
            10.1371/journal.pone.0290097. eCollection 2023"
COMMENT     Sequence Label: pKD008
FEATURES             Location/Qualifiers
     source          1..8748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(208..837)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    2138..2162
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4576..4600
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5033..5848
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5033..5848
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5106..5125)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5936..6524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6425..6444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6678..6695
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6710..6850
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6776..6795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6936..6958
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      7194..7388
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7454..8527)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc ttggtgtgat
       61 cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc aggctggcca
      121 acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt
      181 tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt cagcggccag
      241 cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc
      301 ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc
      361 gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga tcgcccgcga
      421 cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct taaccagctc
      481 caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa tcagcacgaa
      541 gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc cgtcgatcac
      601 tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc
      661 gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg
      721 atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg ctagatgggt
      781 tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa ccttcatgcg
      841 ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac cgcatgacgc
      901 aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt ttcttcagcg
      961 gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag gatttcatgc
     1021 agccgcacgg ttgagaggtg cgcgggcggc tcgaacacgt acccggccgc gatcatctcc
     1081 gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg
     1141 cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt
     1201 cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa
     1261 gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct ttcacggtgc
     1321 ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc
     1381 gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga
     1441 tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg
     1501 ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg ccggcctcct
     1561 ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg
     1621 tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc tccgggcggt
     1681 cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt
     1741 cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag cccagcaggc
     1801 cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa gtattctact
     1861 ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc
     1921 gcgtaactta ggacttgtgc gacatgtcgt tttcagatga cggctgcact gaacgtcaga
     1981 agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc cgaggggaac
     2041 cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa
     2101 atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt
     2161 cagaattacc atggcacctg ctacagtcaa cctcgacgag tcagtaataa acggcgtcaa
     2221 agtggttgca gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc
     2281 tcaacctaaa aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac
     2341 acgtgtcatt ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt
     2401 cctcgtcttt tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca
     2461 attcagattt caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc
     2521 aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc
     2581 caatttcgta tatgttcttt ggtttagatt ctgttaatct tagatcgaag acgattttct
     2641 gggtttgatc gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat
     2701 ttgttcaaat aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag
     2761 atctggtgtt agtttctagt ttgtgcgatc gaatttgtag attaatctga gtttttctga
     2821 ttaacaaaca atggcaccaa aagccggaaa gaaaccagca gagaaaaagc ccgcagagaa
     2881 agctccagca gaagaagaga aagtagccga aaaagcacca gcagagaaga aaccaaaagc
     2941 cggaaaaaaa ctaccaaaag aagcagtcac cggaggagtc gagaagaaga agaaacgagt
     3001 caagaaaagc acagagacat acaagatcta catcttcaag gtgttgaaac aagttcatcc
     3061 agatattgga atctcaagta aagctatggg gattatgaac agttttatca atgatatatt
     3121 cgagaaactt gcgcaagaag cttcaaaatt agctaggtat aataagaaac ctacgattac
     3181 ttctagggag attcagacag ctgttagact agtgttacct ggtgaacttg ctaagcatgc
     3241 tgtttctgaa ggaactaagg ctgttactaa gtttactagc tcttcaggtt cgacagccgc
     3301 tagtgcgtct gcatcaggga gtggttccgg aagcggctct ggatcgggct cagggagtgg
     3361 ttccggcagc ggctctggat cggcggccgc tgcaggctcc gctgctgctt caggttctgg
     3421 ctccggatct ggctcaggta gtgggtccgg atcaggatca ggttcgggtt cagggtccgg
     3481 atctggttcc ggctcttcaa gttcaggtgc tagcggtggc agcggtggca ctagtggtgg
     3541 aggcggatcc atggtgagca agggcgaggc agtgatcaaa gagttcatga ggttcaaggt
     3601 ccacatggaa ggcagcatga acggacatga gttcgagatc gaaggtgaag gtgagggtag
     3661 accttacgag ggaactcaga ctgctaagct caaggtgaca aaaggtggac cgcttccgtt
     3721 cagctgggat attctttcac cgcagttcat gtacggcagc agggctttca tcaagcaccc
     3781 tgctgatatt ccggactact acaagcagtc tttcccagag ggattcaagt gggagcgtgt
     3841 gatgaatttc gaggatggtg gtgctgtgac tgtgacccag gatacttctc ttgaggatgg
     3901 gaccctcatc tacaaggtga agctcagggg aactaacttc cctcctgatg gtcctgtgat
     3961 gcagaaaaag actatgggct gggaagcctc taccgagaga ctttatcctg aggacggtgt
     4021 gcttaagggc gacatcaaga tggctcttag gcttaaggac ggtggacgtt acctcgctga
     4081 tttcaagact acctacaagg ccaagaaacc ggtgcaaatg cctggggctt acaacgttga
     4141 cagaaagctc gacatcacca gccacaacga ggattacact gtggttgagc agtacgagag
     4201 gtctgagggt cgtcattcta ctggtggcat ggacgagctg tacaagtagc tgcgagctca
     4261 tatgaagatg aagatgaaat atttggtgtg tcaaataaat agcttgtgtg cttaagtttg
     4321 tgtttttttc ttggcttgtt gtgttatgaa tttgtggctt tttctaatat taaatgaatg
     4381 taagatctca ttataatgaa taaacaaatg tttctataat ccattgtgaa tgttttgttg
     4441 gatctcttct gcagcatata actactgtat gtgctatggt atggactatg gaatatgatt
     4501 aaagataagg aattcactaa gtgacacatg accataggag aacgtatatc ggcaggtgct
     4561 cgagcaattc gttatgttta caccacaata tatcctgcca ccagccagcc aacagctccc
     4621 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg
     4681 cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga tgctattcgg
     4741 aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt
     4801 tgattgtaac gatgacagag cgttgctgcc tgtgatcacc gcggtttcaa aatcggctcc
     4861 gtcgatacta tgttatacgc caactttgaa aacaactttg aaaaagctgt tttctggtat
     4921 ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg tcttgttata attagcttct
     4981 tggggtatct ttaaatactg tagaaaagag gaaggaaata ataagttctg ttatgagcca
     5041 tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg atgctgattt
     5101 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctaccgatt
     5161 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     5221 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     5281 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg
     5341 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     5401 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     5461 cgatcgcgta tttcgactcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     5521 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     5581 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     5641 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     5701 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     5761 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     5821 tcatttgatg ctcgatgagt ttttctaatc acctagaatg catgaccaaa atcccttaac
     5881 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5941 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6001 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6061 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6601 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6661 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6721 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     6781 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     6841 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6901 cccggcatcc gcttacagac aagctgtgac cgtcaccggg agctgcatgt gtcagaggtt
     6961 ttcaccgtca tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga
     7021 gtggcgacgg cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg ccagccattt
     7081 ttgagcggcc agcggccgcg ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg
     7141 accgaagggt aggcgctttt tgcagctctt cggctgtgcg ctggccagac agttatgcac
     7201 aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc
     7261 cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg
     7321 ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc
     7381 tatccacagg aaagagtcct tttcgacctt tttcccctgc tagggcaatt tgccctagca
     7441 tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca
     7501 tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat
     7561 ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac
     7621 tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg
     7681 ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg
     7741 tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga
     7801 tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg
     7861 cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg
     7921 caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc tgctttccgc
     7981 catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg
     8041 gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca
     8101 tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta
     8161 cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca
     8221 gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca
     8281 tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac
     8341 cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt
     8401 caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact
     8461 tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac
     8521 cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac
     8581 ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc
     8641 ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact
     8701 catttattca tttgctcatt tactctggta gctgcgcgat gtattcag
//
